Cloning and recombinant expression of rat and human kynureninase

FEBS Letters

Nakamura, Masayuki³  Benatti, Luca¹  Cozzi, Liviana²  Gatti, Silvia¹  Okuno, Etsuo³  Breton, Jerome²  Kido, Ryo³  Speciale, Carmela¹  Toné, Shigenobu⁴  Mostardini, Marina¹  Avanzi, Nilla²  Toma, Salvatore² 

[1] CNS Research, Pharmacia and Upjohn, Via Pasteur 10, 20014 Nerviano, Italy;Bioscience Center, Pharmacia and Upjohn, Via Pasteur 10, 20014 Nerviano, Italy;Department of Biochemistry, Wakayama Medical College, 27 Kyubancho, Wakayama 640, Japan;Tokyo Metropolitan Institute of Medical Science, Tokyo 113, Japan

关键词: Kynurenine;    Kynurenic acid;    Excitatory amino acid;    Neuroprotection;    Cysteine-S-conjugate β-lyase;    Quinolinic acid;   

DOI  :  10.1016/S0014-5793(97)00374-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Kynureninase [E.C.3.7.1.3.] is one of the enzymes involved in the biosynthesis of NAD cofactors from tryptophan through the kynurenine pathway. By tryptic and CNBr digestion of purified rat liver kynureninase, we obtained about 28% of the amino acid sequence of the enzyme. The rat kynureninase cDNA, isolated by means of reverse-transcribed polymerase chain reaction and hybridization screening, codes for a polypeptide of 464 amino acids. Northern blot analysis revealed the synthesis of a 2.0 kb rat kynureninase mRNA. A cDNA encoding human liver kynureninase was also isolated. The deduced amino acid sequence is 85% identical to that of the rat protein. COS-1 cells were transfected with both cDNAs. The K _m values of the rat enzyme, for l-kynurenine and dl-3-hydroxykynurenine, were 440±20 μM and 32±5 μM and of the human enzyme 440±20 μM and 49±6 μM, respectively. Interestingly, COS-1 cells transfected with the cDNA coding for rat kynureninase also display cysteine-conjugate β-lyase activity.

【 授权许可】

Unknown